An understanding of the complex interplay between nuclear structure and function requires studies at all levels from molecules up to the entire system. ‘High-throughput’ molecular approaches can explore the average 3D organization of entire genomes within cell populations. They need to be combined with microscopic studies on the single cell level to explore the cell-to-cell variability and to provide a three-dimensional perception of the functional nuclear landscape. Recent developments of super-resolution 3D and 4D light microscopy have revolutionized the power of microscopy (Cremer C, et al. 2011, Biotechnol J 6:1037-1051; Schermelleh L, et al. 2008, Science 320:1332-1336; Huang B, Babcock H, & Zhuang X. 2010, Cell 143:1047-1058) but each microscopic approach has its own advantages and disadvantages.
A biologically meaningful interpretation of high resolution images from nuclei in fixed cells - based on super resolution fluorescence microscopy or electron microscopy - may be tainted both by fixation and imaging artifacts imposed by a given microscopic procedure. Correlative microscopy provides an experimentally demanding but also highly promising approach to better understand these issues. We have developed an approach which allows to view the same cell nucleus first in its living state with wide field or spinning disk laser scanning microscopy and after fixation with confocal light microscopy, super-resolution microscopy (3D structured illumination microscopy, 3D-SIM) and transmission electron microscopy (TEM).
Relevant publications from our group:
Hübner B, Cremer T, Neumann J. Correlative microscopy of individual cells – sequential application of microscopic systems with increasing resolution to study the nuclear landscape. Methods Mol Biol 2013 (in press).
Rouquette J, Cremer C, Cremer T, Fakan S (2010) Functional nuclear architecture studied by microscopy: Present and future. Int Rev Cell Mol Bio 282: 1-156